The overall purpose of this study was to investigate the effects of level of isoflavones supplementation on bone metabolism in growing female rats. The effects of level of isoflavones supplementation on bone mineral density (BMD) and bone mineral content (BMC) were inspected in this study. Forty-five rats divided into three groups: Casein, 1/2IF, IF. The serum and urine concentrations of calcium and phosphorus were determined. BMD and BMC were estimated by using PIXImus (GE Lunar Co, Wisconsin.) in spine and femur on 3, 6, 9 weeks after feeding. This study of results were as follows: The isoflavones supplementation level did not affect weight gain, mean food intake and food efficiency ratio. The serum concentration of calcium, phosphorus were not significantly different by different level of isoflavones supplementation. The urinary calcium and phosphorus excretion were not significantly different, too. Spine and femur BMD, BMC were not significantly increased by different level of isoflavones supplementation on 3 and 6 weeks after feeding. Spine BMD and spine BMC per weight, femur BMC per weight were significantly increased in the groups 1/2IF and IF at the ninth week after feeding, but there was no significant difference by different level of isoflavones supplementation. Spine BMD per weight and femur BMD per weight were significantly higher in the group of IF than in the group of Casein and 1/2IF at the ninth week after feeding. These results suggest that the group of IF with rich isoflavones supplementation was effective to the increase of BMD spine and femur in growing female rats, respectively.
Animals ; Bone Density* ; Calcium ; Caseins ; Eating ; Female* ; Femur ; Humans ; Isoflavones* ; Metabolism ; Phosphorus ; Rats* ; Spine ; Weight Gain

This study was conducted to investigate the effect of soy protein hydrolyzate on lipid metabolism and antioxidant activity in the rat. Thirty-eight male rats of Sprague-Dawley strain were divided into five groups: casein, isolated soy protein (ISP), seoritae protein hydrolyzate (SH), soluble soy protein hydrolyzate (SS), and insoluble soy protein hydrolyzate (IS). The control diet (casein group) contained 20% casein protein and experimental diet contained 10% casein and 10% isolated soy-protein or soy-protein hydrolyzate. Fecal lipid content was increased and lipid apparent absorption rate was decreased significantly by the ISP group at the first week of experimental period. Blood triglyceride, total cholesterol, LDL-cholesterol and atherogenic index (AI) were decreased by soy protein hydrolyzate groups than casein group. Liver total lipid, triglyceride and cholesterol were not different among groups, but showed decreasing tendencies in soyprotein hydrolyzate groups. The lipid lowering effect was prominent in the IS group among soy protein hydrolyzate groups. Total antioxidant activity showed increasing tendency in the seoritae hydrolyzate group. Liver superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase activities also showed higher tendencies in the seoritae hydrolyzate group than other groups. In conclusion, insoluble soyprotein hydrolyzate was more effective in lowering body lipids and seoritae hydrolyzate had higher antioxidant capacity among soy protein hydrolyzates.
Absorption ; Animals ; Caseins ; Catalase ; Cholesterol ; Diet ; Glutathione Peroxidase ; Humans ; Lipid Metabolism ; Liver ; Male ; Rats ; Soybean Proteins ; Sprains and Strains ; Superoxide Dismutase

The study of mammary gland bioreactor is in the ascendant. In order to generate transgenic goats of well-controlled expression of exogenic genes, we constructed a human lactoferrin (hLF) gene targeting vector containing promoter, exon 1, intron1 and some of exon 2 (about 6.1 kb fragment) and exon 6 approximately 9 (about 3.3 kb fragment) of the goat beta-casein gene as well as hLF minigene, neo gene inserted into them and tk gene ligated to the 3' end of the construct. The 9.4 kb goat genomic sequences as homologous arms were initially amplified by PCR with local goat tissue DNA. The expression vector was named pBC-tk-neo-hlf. Then the recombinant plasmid pBC-tk-neo-hlf containing hLF minigene was transfected into mice mammary tumor cell line C127 by liposome, cell clones were selected with G418. After proliferating, the transfected cells were induced with insulin, luteotropic hormone and hydrocortisone. The result of Western-blotting analysis showed that the transfected cells can secrete hLF protein, and the recombinant protein expressed in cultured cell supernatant has the similar molecular weight as the native protein. The expression level detected by ELISA was 0.21 microg/mL. This result indicated that the targeting vector could efficiently direct the expression of hLF in mammary cells,and it confirmed the validity of the constructed vector. At the same time, C127 cell line proved to be useful for evaluating the regulation of a foreign gene expression in mammary gland specific expression vector.
Animals ; Caseins ; genetics ; Cell Line, Tumor ; Goats ; Humans ; Lactoferrin ; genetics ; Mammary Glands, Animal ; cytology ; metabolism ; Mice ; Molecular Weight ; Transfection

This study was carried out to investigate the efficacy of sn-2 palmitic acid-fortified vegetable oil (Sn2PA) on calcium absorption and to confirm the synergistic effects of fructooligosaccharide on calcium absorption. Male SD rats were fed 6 kinds of casein based diets containing vegetable oil (control), sn-2 palmitic acid-fortified vegetable oil (Sn2PA) and Sn2PA with fructooligosaccharide(Sn2PAFO) in two levels of calcium (normal 0.5% and high 1.0%) for 3 weeks. Total lipids, cholesterol, triglyceride and calcium in blood were measured. Feces were collected using cages for 4 days. Serum concentrations of total lipids and calcium were not significantly different among groups. However, serum triglyceride was significantly decreased by fructooligosaccharide supplementation regardless of dietary calcium level. The lipid absorption was not significantly different among experimental groups. Calcium absorption was significantly higher in Sn2PAFO group than other groups. Calcium solubility of intestine was increased by sn-2 palmitic acid supplementation. These results suggest that sn-2 palmitic acid and fructooligosaccharide supplementation could be beneficial for baby foods including infant formula, with regard to increasing absorption of calcium by more soluble calcium in the small intestinal content.
Absorption ; Animals ; Calcium ; Calcium, Dietary ; Caseins ; Cholesterol ; Diet ; Feces ; Gastrointestinal Contents ; Humans ; Infant Formula ; Intestines ; Lipid Metabolism ; Male ; Oligosaccharides ; Palmitic Acid ; Rats ; Solubility ; Vegetables

The effect of the level of casein phosphopeptide (CPP) on mineral (Ca and P) bioavailabilties and bone biomarker of aged ovariectomized (OVX) Sprague-Dawley rats were studied as a model for postmenopausal bone loss. Forty five Spargue dawley rats, 220-230g of body weight were fed a control diet (AIN 93M) or containing different level of CPP diet for 7 weeks: 0% (sham control; SC, OVX control; OC), 1% (OVX low CPP diet; OL), 2% (OVX medium CPP diet; OM), 3% (OVX high CPP diet; OH) Ca absorption was unaffected by increasing CPP content from 0 to 3%. Urinary Ca excretion was increased by OVX, and decreased by CPP significantly (p<0.05) with no evident dose-relationship. The urinary P excretion was increased by CPP intake in OVX rats. The fecal excretion of P given CPP decreased in OVX with dose dependent manner. Ca and P contents of femur significantly increased by adding 2 or 3% of CPP when compared with OC group and OL group (p<0.05). There were no significant differences in serum alkaline phosphatase activity and c-terminal telopeptide excretion in experimental groups. Although ovariectomy induced the increase in urinary c-terminal telopeptide excretion, 2 or 3% of CPP in the diet decreased urinary c-terminal telopetide excretion significantly. These finding suggest the usefulness of CPP in the prevention of postmenopausal bone loss by decreasing urinary Ca excretion and bone resorption. Over 2 percent of CPP in the diet was effective to prevent postmenopausal bone loss.
Absorption ; Alkaline Phosphatase ; Animals ; Body Weight ; Bone Resorption ; Calcium ; Caseins ; Diet ; Female ; Femur ; Humans ; Metabolism* ; Osteoporosis, Postmenopausal ; Ovariectomy ; Phosphorus ; Rats* ; Rats, Sprague-Dawley

This study was performed to evaluate the effect of dietary protein source and sulfur amino acid content on bone metabolism in rats. Thirty male rats (body weight 145 +/- 2 g) were divided into three groups. The rats in the first group were fed on casein 20% diet as animal protein source and those in the second group were fed on soy 20% diet as plant protein source. Sulfur amino acid ratio of these group was 1.07 : 1. The rats in the third group were fed on soy 20% diet and the sulfur amino acid were supplemented with the amount contained as much in the soy 20% diet. All rats were fed on experimental diet and deionized water ad libitum for 9 weeks, The total body, spine, femur bone mineral density and bone mineral content were measured using Dual Energy X-ray Absorptiometry. Calcium, phosphate, pyridinoline, creatinine in urine and calcium, phosphate, alkaline phosphatase, osteocalcin in serum were measured. During the experimental period, plant protein (soy protein) group had a lower urinary Ca excretion, urine pyridinoline & crosslinks value and had a higher Ca efficiency in total bone and femur bone mineral density than animal protein (casein) group. There were no significant differences in serum calcium, phosphate, alkaline phosphatase and osteocalcin among the three groups of the rats. The findings from this study demonstrated that plant protein (soy protein) is beneficial of bone mineral density because it had a higher Ca efficiency in total bone and femur bone mineral density than animal protein (casein). However, the supplementation of sulfur amino acid on soy results were consistent with prior studies that dietary sulfur amino acid load had a negative effect on calcium balance. The rats fed sulfur amino acid supplementation diet increased urinary calcium excretion and decreased calcium efficiency for total and femur mineral density. Therefore, dietary protein source and sulfur amino acid content influence bone metabolism.
Absorptiometry, Photon ; Alkaline Phosphatase ; Animals ; Bone Density ; Calcium ; Caseins ; Creatinine ; Diet ; Dietary Proteins* ; Femur ; Humans ; Male ; Metabolism* ; Osteocalcin ; Plants ; Rats* ; Spine ; Sulfur* ; Water

A simple method of trypsin/collagenase I alternative digestion and iterative culture flask adherence to discard fibroblasts for bovine mammary cell culture was established in this study. By immunohistochemistry, flow cytometry, western blot, Electron microscopy analysis, the characteristics of bovine mammary cells were investigated in vitro. Effect of hyperthermia on the cell ultrastructures was also observed. The results showed that the mammary cells were diploid epithelia with intact 30 pairs chromatins, which could secrete alpha-casein into the medium. After exposed to hyperthermia, the cell condensed chromatin like crescent on the nuclei verges, mitochondria occurred expansion and vacuolization, and apoptotic bodies appeared, which suggested that heat stress could induce apoptosis of the mammary epithelia.
Animals ; Apoptosis ; Blotting, Western ; Caseins ; metabolism ; Cattle ; Cell Line ; Cell Nucleus ; ultrastructure ; Cells, Cultured ; Chromatin ; ultrastructure ; Epithelial Cells ; cytology ; metabolism ; ultrastructure ; Female ; Flow Cytometry ; Hot Temperature ; Immunohistochemistry ; Mammary Glands, Animal ; cytology ; metabolism ; Microscopy, Electron, Transmission ; Mitochondria ; ultrastructure ; Vimentin ; metabolism

Whey protein concentrate (WPC 80) and sodium caseinate were hydrolyzed by Protamex to 5%, 10%, 15%, and 20% degree of hydrolysis (DH). WPC 80, sodium caseinate and their hydrolysates were then analyzed, compared and evaluated for their nutritional qualities. Their chemical composition, protein solubility, amino acid composition, essential amino acid index (EAA index), biological value (BV), nutritional index (NI), chemical score, enzymic protein efficiency ratio (E-PER) and in vitro protein digestibility (IVPD) were determined. The results indicated that the enzymatic hydrolysis of WPC 80 and sodium caseinate by Protamex improved the solubility and IVPD of their hydrolysates. WPC 80, sodium caseinate and their hydrolysates were high-quality proteins and had a surplus of essential amino acids compared with the FAO/WHO/UNU (1985) reference standard. The nutritive value of WPC 80 and its hydrolysates was superior to that of sodium caseinate and its hydrolysates as indicated by some nutritional parameters such as the amino acid composition, chemical score, EAA index and predicted BV. However, the E-PER was lower for the WPC hydrolysates as compared to unhydrolyzed WPC 80 but sodium caseinate and its hydrolysates did not differ significantly. The nutritional qualities of WPC 80, sodium caseinate and their hydrolysates were good and make them appropriate for food formulations or as nutritional supplements.
Amino Acids ; chemistry ; Caseins ; chemistry ; metabolism ; Dietary Proteins ; analysis ; Evaluation Studies as Topic ; Hydrolysis ; Milk Proteins ; chemistry ; metabolism ; Models, Statistical ; Nutritive Value ; Protein Hydrolysates ; chemistry ; Solubility ; Temperature ; Time Factors ; Tryptophan ; chemistry ; Whey Proteins

OBJECTIVETo establish an appropriate neonatal rat model of necrotizing enterocolitis (NEC) and to investigate the protective effects of glycomacropeptide (GMP) on the gut from injury in neonatal rats with NEC.

METHODA total of 36 neonatal SD rats were randomly divided into 3 groups: NEC model group (Group M), NEC + GMP group (Group G) and normal control group (Group N), each group had 12 rats. All the neonatal rats were fed with breast milk in the first 3 days after birth. During the second 3 days after birth, the rats of Group N were still maternal breast-fed, but the rats of Group M and Group G were separated from their mothers and lived in incubator and began to be formula fed, and were subjected to cold exposure shortly after hypoxic-reoxygenation treatment. After being fed in such means for 6 days, all the neonatal rats were placed into the incubator and fasted for 24 hours. Then all the rats were sacrificed by cervical dislocation. Intestinal tissue located at the boundary of ileum and cecum was obtained for: (1) histological examination after HE staining, (2) TUNEL detection, (3) electron microscopic observation; and the tissue homogenate was obtained for checking TNF-α and IL-1β levels by ELISA and platelet activating factor (PAF) mRNA expression by quantitative fluorescence (QF)-PCR.

RESULT(1) The pathological scores of the 3 groups were 2.17 ± 0.83 (Group M), 0.92 ± 0.79 (Group G) and 0.17 ± 0.39 (Group N) separately. There was significant difference between Group M and Group G (H = 8.819, P = 0.003). (2) TNF-α levels of 3 groups were (41.94 ± 13.51) pg/ml (Group M), (31.69 ± 11.68) pg/ml (Group G) and (17.42 ± 7.18) pg/ml (Group N) separately, and TNF-α level in Group G was significantly lower than that of Group M (F = 3.959, P = 0.030). (3) IL-1β levels of 3 groups were (150.33 ± 36.41) pg/ml (Group M), (118.36 ± 33.00) pg/ml (Group G) and (28.44 ± 15.04) pg/ml (Group N) separately, and IL-1β level in Group G was lower than that of Group M (F = 5.080, P = 0.013). (4) Expression levels of intestinal PAF mRNA (2(-ΔΔCt) value): 3.01 ± 0.96 (Group M), 1.56 ± 0.29 (Group G), 1.01 ± 0.13 (Group N), the level of Group G was significantly lower than that of Group M (F = 25.251, P = 0.000). (5)Electron microscopy: Group N showed that its cell volume was mostly occupied by the nucleus, the structure was clear, nuclear membrane existed, suggesting the normal phase of cell; Group M showed that apoptotic body existed, suggesting that the advanced stage phase of apoptosis; Group G showed that condensed chromatin marginated around the nuclear envelope, nuclear pores expanded, suggesting the early phase of apoptosis. (6) The apoptosis rate of intestinal epithelial cells by TUNEL detection: 38.79 ± 9.79 (Group M), 29.54 ± 7.30 (Group G), 6.37 ± 1.96 (Group N); the apoptosis rate of intestinal epithelial cells of Group G was significantly lower than that of Group M (F = 6.888, P = 0.003).

CONCLUSIONGMP has protective effects on guts of neonatal rats with NEC, which may probably work by reducing TNF-α, IL-1β and PAF expression, inhibiting the apoptosis of intestinal epithelial cells and reducing intestinal tissue injury.

Animals ; Animals, Newborn ; Apoptosis ; Caseins ; pharmacology ; Cold Temperature ; Enterocolitis, Necrotizing ; drug therapy ; metabolism ; pathology ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Female ; Hypoxia ; complications ; Interleukin-1beta ; metabolism ; Intestinal Mucosa ; metabolism ; pathology ; Intestines ; metabolism ; pathology ; Male ; Peptide Fragments ; pharmacology ; Platelet Activating Factor ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

BACKGROUND/OBJECTIVES: Soy isoflavones are structurally similar to estrogen and bind to estrogen receptors, suggesting that they exhibit estrogenic activities; therefore, they are referred to as phytoestrogens. Fermentation may affect the bioavailability of isoflavones altering soy isoflavone glycosides in the form of aglycones. Thus, this study investigated the effects of fermented soybeans by Rhizopus oligosporus on bone metabolism in both young rats as a pilot test and in ovariectomized (ovx) old rats as a model of menopause. MATERIALS/METHODS: In the pilot test, a total of 24 seven-week-old female Sprague-Dawley (SD) rats were fed one of three diets for a period of four weeks: casein, unfermented soybean product, or fermented soybean product by R. oligosporus. In the ovx rat model, 20-week-old SD rats weighing 260-290 g underwent either sham-operation (n = 10) or bilateral ovariectomy (n = 30) and were then fed the AIN-93M diet for one week. Thereafter, rats were fed sham-casein, ovx-casein, ovx-soybean, or ovx-fermented soybean diet for five weeks. After decapitation, femoral bones were isolated and preserved in 9% formalin for assessment of bone mineral density (BMD), bone mineral content (BMC), and bone-breaking strength (BBS). RESULTS: Ovx rats showed significantly increased weight gain and decreased uterine wet weight. Of particular interest, ovx rats fed fermented soybeans showed increased uterine wet weights compared to control rats. Fermented soybean diet caused a significant increase in plasma 17-beta estradiol concentrations in young rats, and 17-beta estradiol levels were enhanced in ovx rats to match those of sham-operated ones. Significantly lower femoral BMD and BMC were observed in ovx rats compared to sham-operated controls, whereas bone areas did not differ statistically among the groups. In addition, BBS tended to be increased in ovx rats fed soybeans and fermented soybeans. CONCLUSIONS: Supplementation of fermented soybeans could have preventive and therapeutic effects against osteoporosis in postmenopausal women.
Animals ; Biological Availability ; Bone Density ; Caseins ; Decapitation ; Diet ; Estradiol ; Estrogens ; Female ; Fermentation ; Formaldehyde ; Glycosides ; Humans ; Isoflavones ; Menopause ; Metabolism ; Models, Animal ; Osteoporosis ; Ovariectomy ; Phytoestrogens ; Plasma ; Rats* ; Rats, Sprague-Dawley ; Receptors, Estrogen ; Rhizopus* ; Soybeans* ; Weight Gain ; Weights and Measures